Strategies for Measuring Damage and Repair in Gene-Sized Specific DNA Sequences
Interest in how the efficiency of DNA repair might vary among specific categories of cellular DNA dates almost to the origin of the “repair replication” technique, which quantifies the short stretches of DNA synthesized during excision repair (Pettijohn and Hanawalt, 1964). It has always been clear that the biological consequences of DNA damage to the cell or organism would depend strongly on the functional role of the particular segment of DNA suffering the damage. Early studies were confined to comparing repair in classes of DNA that were in relative abundance and could be physically isolated for analysis such as chloroplast and mitochondrial DNA, and genomic satellite DNA. Later, the repair of the highly repetitive alpha DNA sequences in African green monkey cells was investigated in detail using a variety of techniques. This was made possible by the abundance of this alpha DNA species; 8% of the DNA can be easily isolated as pure 172-base-pair fragments by digestion by HindIII and gel electrophoresis (Zolan et al., 1982). These investigations (reviewed in Smith, 1987) demonstrated complex differences in the repair of this nontranscribed sequence as compared to the remaining, bulk DNA, and gave impetus to efforts to develop methods for studying repair in active genes.
KeywordsRepair Patch Cyclobutane Pyrimidine Dimer DHFR Gene Lesion Frequency Adenine Phosphoribosyltransferase
Unable to display preview. Download preview PDF.
- Baird, W. M., Smith, C. A., Spivak, G., Mauthe, R. J., and Hanawalt, P. C. (1994). Analysis of the fine structure of the repair of anti-benzo[a]pyrene-7,8-diol-9,10-epoxide-DNA adducts in mammalian cells by laser-induced strand cleavage. Polycyclic Aromatic Compounds 6:169–176.CrossRefGoogle Scholar
- Bohr, V. A., and Okumoto, D. S. (1988). Analysis of pyrimidine dimer repair in defined genes, in:DNA Repair: A Laboratory Manual of Research Procedures, Volume III (E. C. Friedberg and P. C. Hanawalt, eds.), Dekker, New York, pp. 347–366.Google Scholar
- Leadon, S. A. (1988). Immunological probes for lesions and repair patches in DNA, in:DNA Repair: A Laboratory Manual of Research Procedures, Volume III (E. C. Friedberg and P. C. Hanawalt, eds.), Dekker, New York, pp. 311–326.Google Scholar
- Smith, C. A. (1988). Repair of DNA containing furocoumarin adducts, in:Psoralen DNA Photobiology, Volume II (F. Gasparro, ed.), CRC Press, Boca Raton, FL, pp. 87–116.Google Scholar
- Spivak, G., and Hanawalt, P. C. (1995). Determination of damage and repair in specific DNA sequences, in:Methods: A Companion to Methods in Enzymology, Vol. 7, Academic Press, London, pp. 147–161.Google Scholar
- van Hoffen, A., Venema, J., Meschini, R., van Zeeland, A. A., and Mullenders, L. H. (1995). Transcription-coupled repair removes both cyclobutane pyrimidine dimers and 6–4 photoproducts with equal efficiency and in a sequential way from transcribed DNA in xeroderma pigmentosum group C fibroblasts. EMBO J. 14:360–367.PubMedGoogle Scholar
- Vos, J.-M. (1988). Analysis of psoralen monoadducts and interstrand crosslinks in defined genomic sequences, in:DNA Repair: A Laboratory Manual of Research Procedures, Volume III (E. C. Friedberg and P. C. Hanawalt, eds.), Dekker, New York, pp. 367–398.Google Scholar